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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE
KARNATAKA
ANNEXURE−II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
NAME OF THE CANDIDATE Dr. DISHA NAGPAL
AND ADDRESS (IN BLOCK POST GRADUATE STUDENT,
LETTERS)
DEPARTMENT OF PERIODONTICS,
COLLEGE OF DENTAL SCIENCES,
DAVANGERE−577 004
KARNATAKA.
2.
NAME OF THE
COLLEGE OF DENTAL SCIENCES,
INSTITUTION
DAVANGERE−577 004
KARNATAKA.
3.
COURSE OF STUDY AND
MASTER OF DENTAL SURGERY IN PERIODONTICS
SUBJECT
4.
DATE OF ADMISSION TO 22nd MAY 2012
COURSE
5.
TITLE OF THE TOPIC
DETECTION
AND
COMPARISON
OF
SELENOMONAS SPUTIGENA IN SUBGINGIVAL
BIOFILMS
IN
CHRONIC
PERIODONTITIS PATIENTS.
AND
AGGRESSIVE
6.
BRIEF RESUME OF THE INTENDED WORK :
6.1 Need for the study :
The role of oral microbiota in etiology of periodontal disease has been well established and
specificity may exist among certain bacterial species/groups and the various forms of periodontal
disease.1 The complexity and diversity of periodontal microbiota has been confirmed by numerous
studies.2 However, only a few species have been recognised as periodontal pathogens, namely
Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia.3
The microbiota of localized and generalised forms of aggressive periodontitis contain higher
proportions of Aggregatibacter actinomycetemcomitans compared with that of chronic
periodontitis, whereas the proportions of red complex pathogens,4 do not differ between these
periodontal conditions, despite their clinical differences.5 These observations suggest that further
analyses of the microbiota are necessary in order to explain differences in clinical outcomes.It was
observed that Selenomonas species dominated the disease sites of subjects with Generalized
Aggressive Periodontitis6
and Chronic Periodontitis.2 Selenomonas sputigena was the most
frequently detected bacterial species which was multiflagellated , motile, anaerobic rods and
further characterised as etiological agent in periodontal disease.7
However, the detection of this bacteria is needed because its etiological and pathological role in
periodontal diseases is still unclear and information on absolute numbers and proportions of
organisms in samples is important in distinguishing the species associated with periodontal
health/disease and to evaluate the effects of periodontal therapy.
Therefore, the purpose of the present study is to detect and compare the levels of Selenomonas
sputigena in subgingival biofilms from patients with Aggressive Periodontitis, Chronic
Periodontitis and Periodontally healthy control subjects.
.
6.2 Review of literature :
A study was conducted to detect oral motile bacteria directly from dental plaque, specific PCR
primers for Centipeda periodontii and Selenomonas sputigena were designed based on the
sequence analysis of their 16S rDNA. The primers were specific and sensitive enough to amplify
DNA fragments from the available oral bacteria. The detection limit was fewer than 10 bacterial
cells per sample. It was also possible to detect these bacteria in dental plaque. The prevalence of
these bacteria varied in each sample. The specific primers designed in this study clarified the
epidemiology of periodontal disease.8
A study was conducted to compare the levels of Selenomonas sputigena and uncultivated/unrecognized
Selenomonas species in subgingival biofilms from periodontally healthy subjects and from subjects with
Generalized Aggressive Periodontitis. The results showed that S. sputigena and Mitsuokella sp. Human
Oral Taxon (HOT) 131 may be associated with the pathogenesis of generalized aggressive periodontitis, and
their role in the onset and progression of this infection should be investigated further.9
The microbial population in 73 Rapidly Progressive Periodontitis (RPP) lesions in 10 young adults aged 2535 years(5 males,5 females)was studied in relation to the clinical parameters probing depth, bleeding on
probing and suppuration, which were recorded at the sampled sites. Significant differences between
bleeding index 0, 1, and 2 (P < 0.05) in frequency of detection were found for P. intermedia, Campylobacter
concisus, Selenomonas sputigena, and Peptostreptococcus micros at bleeding sites and for Streptococcus
sanguis, Actinobacillus actinomycetemcomitans, and B. forsythus (P<0.001) at non-bleeding sites.10
In yet another study, clinical, microbiological and immunological factors were examined using data
from a subject with periodontosis. The subject was monitored at bimonthly intervals for 26 months
at 6 sites per tooth for redness, plaque, suppuration, bleeding on probing, pocket depth, and
attachment level. Subgingival plaque samples were taken from these sites for predominant
cultivable and dark field evaluation before, 5 months and 13 months after treatment by Widman
flap surgery and systemic tetracycline. The proportions of Actinobacillus actinomycetemcomitans
and Selenomonas sputigena were elevated in active sites, while proportions of Bacteroides
intermedius were elevated in control sites. 5 months after treatment, proportions of A.
actinomycetemcomitans, S. sputigena and Eikenella corrodens were significantly decreased in the
previously active sites and proportions of B. intermedius and E. corrodens were significantly
decreased in the control sites.11
A study was conducted to quantitatively compare the bacterial population structure in plaque from
the gingival margin of two groups of patients – one with gingivitis and another with necrotizing
ulcerative
gingivitis
nucleatum/Fusobacterium
(NUG).The analyses showed that the fusiform taxa (Fusobacterium
periodonticum,
Leptotrichia
buccalis,
Tannerella
forsythensis,
and
Capnocytophaga sp.), Campylobacter rectus, Prevotella intermedia, Prevotella nigrescens, Selenomonas
sputigena and treponemes were present in both groups with high prevalence. Porphyromonas gingivalis and
Actinomyces gerencseriae were much more prevalent in the NUG group.12
6.3 Objectives of the study :
1 )To detect the levels of Selenomonas sputigena in subgingival biofilms from patients with
Aggressive Periodontitis,Chronic Periodontitis and Periodontally healthy control subjects.
.
2)To compare the levels of Selenomonas sputigena in subgingival biofilms from patients
with Aggressive Periodontitis,Chronic Periodontitis and Periodontally healthy control
subjects.
7.MATERIALS AND METHODS :
7.1 Source of data :
The patients for this study will be selected from the out patient Department of Periodontics,
College Of Dental Sciences, Davangere, Karnataka. The subjects between 20-55 years of age will
be included in this study.
7.2 Method of Collection of Data (including sampling procedure, if any) :
Sample size: 90 patients ( 45 males and 45 females).
Study design: A clinical and microbiological study with a total of 90 patients (45 males,45
females) with Chronic Periodontitis, Generalized Aggressive Periodontitis and Periodontally
healthy subjects will be included in this study. The patients are divided into 3 groups as follows:
Group I (30) - Periodontally healthy subjects
Group II (30) - Chronic Periodontitis Subjects
Group III (30) - Generalized Aggressive Periodontitis Subjects.
Study period :
The duration of this clinical study will be one year, from January 2013-January 2014.
Clinical parameters :
Following clinical parameters will be recorded at baseline visit using a UNC15 probe:
1. Plaque Index (Silness P. and Loe H., 1964)
2. Gingival Index (Loe H. and Silness J, 1963)
3. Gingival bleeding index (Ainamo & Bay, 1975)
4. Probing Pocket Depth
5. Clinical Attachment Level
6. Suppuration[Present(1) or Absent(0)]
7. Radiographic assessment( Orthopantomogram )
SELECTION CRITERIA:
7
S These patients will be selected consecutively during the study period as and when they are present
with following inclusion and exclusion criteria from both the sexes. The medical and dental
histories will be obtained and a full mouth periodontal examination will be performed. Based on
these data, periodontal diagnosis will be made and subjects who will fulfill the inclusion and
exclusion criteria will be participating in this study.
Inclusion criteria:
Subjects had to have at least 20 teeth and needed to meet the following criteria in order to be
included in this study:
Periodontally healthy9 —
1)20-55 years(15 males and 15 females)
2) Sites with probing depth and clinical attachment level measurements of ≤ 3 mm.
3) ≤ 10% of sites exhibiting bleeding on probing.
Generalized Aggressive Periodontitis9 —
1) ≤ 30 years of age (15 males and 15 females)
2) A minimum of six permanent incisors and/or first molars with at least one site each with probing
depth and clinical attachment level of ≥ 5 mm with vertical bone loss radiographically.
3) A minimum of six teeth (other than first molars and incisors) with at least one site each with
probing depth and clinical attachment level of ≥ 5 mm with vertical bone loss radiographically.
4)Familial aggregation (at least one other member of the family presenting, or with a history of
periodontal disease)
Chronic Periodontitis13 –
1)Age>35 years(15 males and 15 females)
2) >30% of the sites involved with periodontal destruction.
3) ≥5mm of Clinical Attachment Loss and Probing Pocket Depth with vertical/horizontal bone loss
radiographically.
Exclusion criteria:
1) Pregnant or lactating females.
2) Subjects who are smokers.
3) No systemic condition that might influence periodontal status. ( e.g. diabetes, immunological
disorders etc.)
4) Patients on antibiotics, steroids, contraceptive drugs/periodontal therapy in preceding 6
months/during the study.
Microbiological Examination:
Sample collection—The microbial plaque samples were taken prior to the clinical measurements
after removing the supragingival plaque at baseline visit, from each selected patient the pooled
plaque samples were collected with paper points from 3 interproximal sites with >5mm of clinical
attachment loss by using Modified Slot Technique10 for 30 seconds.
The collected samples will be subjected for microbiological analysis i.e. detection and comparison
of Selenomonas sputigena by using 16S rDNA based PCR assay.
Statistical Analysis :
Results obtained will be subjected for appropriate statistical analysis.
Test for proportions (Z test) will be used to compare between groups.
Categorical data will be analyzed by Chi square test.
7.3 Does the study require any investigations or interventions to be conducted on patients or
other humans or animals? If so, please describe briefly.
Yes, this study involves the collection of the subgingival plaque samples from the Aggressive
Periodontitis, Chronic Periodontitis and Periodontally healthy subjects.
7.4 Has ethical clearance been obtained from your institution in case of 7.3?
Yes
8.
REFERENCES:
1. Albandar JM, Brown LJ, Löe H. Putative periodontal pathogens in subgingival plaque of
young adults with and without early-onset periodontitis. J Periodontol 1997;68(10):973–81.
2. Paster BJ, Boches SK, Galvin JL et al. Bacterial diversity in human subgingival plaque.
J Bacteriol 2001;183(12):3770–83.
3. American Academy of Periodontology. Consensus report. Periodontal diseases: pathogenesis
and microbial factors. Ann Periodontol 1996;1:926–32.
4. Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr. Microbial complexes in
subgingival plaque. J Clin Periodontol 1998;25(2):134–44.
5. Faveri M et al Microbiological profile of untreated subjects with localized aggressive
periodontitis. J Clin Periodontol 2009;36(9):739–49.
6. Faveri M et al Microbiological diversity of generalized aggressive periodontitis by 16S rRNA
clonal analysis. Oral Microbiol Immunol 2008;23(2):112–8.
7. Kokeguchi S, Tsutsui O, Kato K, Matsumura T. Isolation and characterization of
lipopolysaccharide from Centipeda periodontii ATCC 35019. Oral Microbiol Immunol
1990;5(2):108-12.
8. Sawada S, Kokeguchi S, Takashiba S, Murayama Y. Development of 16S rDNA-based PCR
assay for detecting Centipeda periodontii and Selenomonas sputigena. Lett Appl Microbiol
2000;30(6):423-6.
9. Gonçalves LF et al Levels of Selenomonas species in generalized aggressive periodonitis. J
Periodont Res 2012; 47(6): 711–8.
10. Kamma JJ, Nakou M, Manti FA. Microbiota of rapidly progressive periodontitis lesions in
association with clinical parameters. J Periodontol 1994;65(11):1073-8.
11. Haffajee AD, Socransky SS, Ebersole JL, Smith DJ. Clinical, microbiological and
immunological features associated with the treatment of active periodontosis lesions. J Clin
Periodontol 1984;11(9):600-18
12. Gmür R, Wyss C, Xue Y, Thurnheer T, Guggenheim B. Gingival crevice microbiota from
Chinese patients with gingivitis or necrotizing ulcerative gingivitis. Eur J Oral Sci 2004;112(1):
33–41.
13. Newman MG,Takei HH,Klokkevold PR,Carranza FA. Carranza’s Clinical Periodontology.
10th edn.Noida,India.2009. p.106.
9.
SIGNATURE OF CANDIDATE
10.
REMARKS OF THE GUIDE
11.
NAME & DESIGNATION OF
Dr. SHOBHA PRAKASH M.D.S.
(IN BLOCK LETTERS)
PROFESSOR AND HEAD,
11.1 GUIDE
DEPARTMENT OF PERIODONTICS,
COLLEGE OF DENTAL SCIENCES,
DAVANGERE-577 OO4.
11.2 SIGNATURE
11.3 CO-GUIDE (IF ANY)
−
11.4 SIGNATURE
−
11.5 HEAD OF THE
DEPARTMENT
Dr. SHOBHA PRAKASH M.D.S.
PROFESSOR AND HEAD,
DEPARTMENT OF PERIODONTICS,
COLLEGE OF DENTAL SCIENCES,
DAVANGERE-577 OO4.
11.6 SIGNATURE
12.
12.1 REMARKS OF THE
CHAIRMAN
PRINCIPAL
&
Dr. V. V. SUBBA REDDY M.D.S.
PRINCIPAL,
COLLEGE OF DENTAL SCIENCES,
DAVANGERE-577 004.
12.2 SIGNATURE